Compositions and methods for repelling coleopterans

ABSTRACT

Compositions and methods for repelling coleopterans. The compositions contain a substrate and an anti-aggregation pheremone intermixed within the substrate. The pheremone may be verbenone, methyl salycylate, wintergreen oil, or combinations thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/773,879, filed on Jan. 27, 2020, which was a continuation of U.S.patent application Ser. No. 15/594,258, filed on May 12, 2017, which wasa continuation-in-part of U.S. patent application Ser. No. 14/938,701filed on Nov. 11, 2015, which also claims the benefit of U.S.Provisional Application No. 62/078,081, filed on Nov. 11, 2014, and theteachings of which are expressly incorporated by reference.

STATEMENT RE: FEDERALLY SPONSORED RESEARCH/DEVELOPMENT

Not Applicable

BACKGROUND 1. Field of the Invention

The present invention relates to products, systems, and methods forcontrolling coleopteran, including, among others, Hypothenemus hampei(coffee berry borer, CBB), the most economically important pest ofcoffee worldwide. More specifically, this invention relates to methodsand systems for repelling coleopteran, for purposes of both populationcontrol and crop protection through the use of volatile semiochemicalswith potent and varied behavioral effects. This semiochemicalformulation can be employed in a broad range of means, including amonolithic repellent and repellent-impregnated rubber to be deployed incontrolled areas, and a liquid formulation that can be amenable to sprayapplication.

2. Background of the Invention

Chemicals existing within the environment that convey some form ofinformation, known as semiochemicals, are used extensively by arthropodsto communicate with each other and can be used in strategies for pestmanagement.

The direct management of insect pests using semiochemicals for repellentapproaches can provide excellent suppression of key coleopteran pests inagriculture and forestry. Large-scale implementation projects haveyielded significant reductions in pesticide use while maintainingacceptably low crop-damage levels.

The coffee berry borer (CBB), Hypothenemus hampei, is the mosteconomically significant pest of coffee worldwide, creating an estimated$500 million in losses annually. The beetle is native to Angola, howeverit extended its range throughout Africa during the early 20th centuryand has continued to do so since. It is believed that most invasionsoccur due to the importation of contaminated seeds, colloquially beans.Regardless of the mode of invasion, CBB has effectively extended itsrange to every major coffee producing country in the world, makinglandfall in the U.S. state of Hawaii in 2010.

The coffee industry has an estimated global economic value between $70and $90 billion annually. Per pound retail prices for coffee can varydrastically depending on a variety of product parameters and theconsumer market, for example per pound coffee prices within the U.S.market range from $2-$30 depending on the product. Due to these pricingvariations the economic impact of CBB losses to some cultivars issubstantially greater than others. When densities of CBB reach very highlevels they can cause yield losses upwards of 35% with 100% of berriesinfested at harvest time, greater losses can occur if harvest isdelayed. In addition to the direct loses, CBB damage can devalue coffeeprices by 30-40%.

The CBB is such a destructive force in the coffee industry because theentire larval stage, and most of the adult stage, is spent within thecoffee berry. Males never emerge from the berry, therefore the entirelife cycle of a male CBB from development to mating is confined to thesame coffee berry it hatched from. Female CBBs, on the other hand,remain within the coffee berry up until mating. Once mated, female CBBsattack immature and mature coffee berries. The attacking female bores ahole in the coffee berry and lays eggs amongst the seeds at a rate of 2to 3 eggs per day for a period of 20 days. The larvae feed upon theseeds until they mature and mate, repeating the cycle. There is a 10:1sex ratio favoring females; allowing for rapid proliferation throughouta coffee plantation. Further, as most of the life cycle occurs insidethe berries, control strategies, both chemical and non-chemical, aregreatly limited.

Because female CBB are able to fly and their lifespan allows for longdurations of activity, (nearly six months under the correctenvironmental conditions) damage to developing berries can occur yearround. Three types of damage have been reported due to CBB: 1) prematurefall of young berries, 2) increased vulnerability of infested ripeberries to secondary infection, and 3) reduction in both yield andquality of coffee, reducing the profitability of the crop.

Historically, coffee farmers have relied on Endosulfan for the controlof CBB, but as of April 2011, it has been phased out globally.Endosulfan is a highly toxic chemical that poses significant risk tohuman health by acting like an endocrine disruptor. It also poses highpotential for bioaccumulation, which can negatively impact entireecosystems surrounding coffee plantations. Further, CBB's life cyclemakes it nearly inaccessible to most control measures. Coffee farmershave been left in an extremely difficult position where they must combatCBB with insecticides that have not yet been evaluated for effectivenessagainst CBB or labeled for use in coffee. Most insecticides used to datehave proven ineffective for control of CBB.

Mycoinsecticide control products containing the spores of the fungusBeauveria bassiana, such as the EPA registered BOTANIGARD® ES (BioWorks,Victor, N.Y., US), have demonstrated the ability to reduce femaleoviposition rates and larvae production and increase CBB mortality.However, the product requires two separate applications, one mid and onelate season, directly to the berries and surrounding ground cover,increasing planning and logistical requirements. Additionally, theproduct costs $60-$90 per acre per application, making overallprotection costly.

Alternatively, coffee farmers can employ various cultural control orhygiene approaches to reduce CBB populations within the plantation:removal of berries and trapping prior to and during fruit development,contained harvesting, and post harvest plantation sanitation.

Pruning fruit-producing trees is a necessity in maintaining a healthyand effective growth operation. Two methods of pruning exist withincoffee farming: 1) pruning one or two verticals on the tree each year(the Kona method), or 2) pruning all the verticals on the tree in thesame year every three to five years (the Beaumont-Fukunaga method).However, prior to pruning, any berries on the trees must be removed toensure that they do not fall to the ground. Research has indicatedberries not removed before pruning can result in as many as 3.2 millionimmature and adult CBBs per acre. Further, the CBBs within the berriesremained reproductively active and continued to emerge for up to threemonths after being on the ground. In order to minimize the risk ofexisting populations continuing into the next season, all berriesremoved prior to pruning must either be buried or burned.

Coffee has a prolonged and variable fruiting season, flowering dependson several environmental conditions, berry growth is slow, and allberries do not ripen simultaneously. This poses an on-going challengefor coffee farmers attempting to intercept female CBBs as they emergefrom their birth berry and seek out a new, uninhabited host berry.Traps, such as the BROCAP® (CIRAD, Paris, FR), can be installed at arate of 8 per acre and suspended from the coffee tree at 1.5 m. Trapsnecessitate a lure composed of methanol and ethanol mixed to variousratios. Baited traps do not guarantee the removal of the CBB from thefield, although at sufficient trap density protection can be achieved.More often traps are used in an effort to monitor for the presence ofthe beetle in the field.

During harvest, emphasis is placed on removing all ripe and droppedfruit. Harvest bags should be tied shut at harvest to avoid the escapeand dispersal of CBB. The processing of berries to seeds should beperformed in closed or semi-closed structures to prevent the escape ofCBBs and allow for proper disposal. Additionally, during post harvest,dried and dropped berries should be collected from the field and buriedor burned as they were at the beginning of the season.

There may exist alternative and/or more effective methods to controlCBB. CBB, like most beetles, uses chemical signaling systems todetermine many aspects of its life cycle. Semiochemicals, artificiallyor naturally sourced, can be used to influence these chemical pathways.One common function of these semiochemicals in beetle populations isrepellency; communicating to incoming beetles that a particular host isoverpopulated and no longer suitable for colonization.

CBB is a bark beetle and member of the sub-family Scolytidae. Thissub-family includes a genus of beetles, Dendroctonus, that are wellknown major pests to global timber industries and as such have wellresearched and documented semiochemical communication systems. Onesemiochemical, verbenone, has shown to be strongly repellent to theDendroctonus genus and may be strongly repellent to the entireScolytidae sub-family.

Prior studies have shown that on average, CBB chose to travel down apath not containing verbenone. In these experiments a Y-tubeolfactometer was used to force CBB to travel one of two paths in orderto elucidate what impact, if any, specific semiochemicals, known toinfluence the behavior of other bark beetles, have on CBB. The resultsconfirmed that verbenone is unattractive to CBB and can potentially becorrelated, but not confirmed, to have a repellant effect. However, thestudy did not provide any specific information on the dosage orverbenone type used.

What was not previously known, and has been now discovered, is thatexposure to (1S)-(−)-Verbenone (cas #1196-01-6), as well as otherverbenone enantiomers will cause both male and female CBBs to dislodgefrom the coffee berry. The invention herein pertains to use of allpotential verbenone sources, including synthetic sources, naturalsources or any natural extract containing verbenone for the use ofrepelling or otherwise modifying the behavior of Hypothenemus hampei,the coffee berry borer. The present discovery applies specifically tothe amount of verbenone, including all verbenone enantiomers, requiredto achieve such repellency. Applications of 1-10,000 g of verbenone(Active Ingredient, A.I.) per acre, using wax emulsions or any otherfield application technology, will be sufficient to repel CBB from acoffee field.

Verbenone has been discovered to be an efficacious repellant to CBBwithin a coffee plantation. The repellent effect will be used to pushCBB from a currently infested plantation, decreasing or potentiallyremoving the infestation within the application area. The technologydisclosed herein may also be used to repel CBB and prevent itsestablishment in low or non-infested plantations, such as new plantings,aggressively controlled areas or geographic regions where CBB has notentered.

Another aspect of the presently disclosed invention is the use ofverbenone as an accessibility tool. By repelling CBB from the coffeeberries, CBB becomes accessible to insecticide sprays. Such an increasein accessibility will lead to more effective insecticide regimes and toa more efficient use of such insecticides.

It is believed that that 1-2,500 verbenone point sources per acre, usingmonolithic or any other field application technology, will provideeffective repellency to CBB.

Based on the considerations above, verbenone acts as a suitable CBBrepellent when formulated with the Specialized Pheromone and LureApplication Technology (SPLAT) described in U.S. Pat. No. 7,887,828, theentirety of which is incorporated by reference herein. This formulationfor Coffee Berry Borer of the present invention (coined SPLAT Verb, andused as such herein) has a strong repellent effect on adult CBB, at low,economically viable doses; it is produced commercially and in largequantities, is labeled organic and registered for “all crops”; and it isa stable molecule (once UV-protected and anchored by SPLAT).

BRIEF SUMMARY

One embodiment of the present disclosure is directed toward systems andmethods for controlling arthropod populations at the adult stage. Thesystems and methods of the present invention utilize repellents that arereactive upon an adult stage arthropod.

One embodiment of the present invention is directed toward a repellentsystem for controlling arthropod populations. The arthropods arecharacterized by a semiochemical-affected adult stage. The system ofthis embodiment includes a polymeric substrate and a semiochemical. Thesemiochemical is reactive upon the adult stage arthropod and isintermixed within the polymeric substrate. The semiochemical may be apheromone, and may specifically be an anti-aggregation pheromone thatdisrupts the behavior of the adult arthropod.

The polymeric substrate may be present in numerous differentconfigurations, including, but not limited to, microspheres, latexsolutions, hot melt glues, resins, plastic flakes, and wax emulsions.The waxes that may be used in the wax emulsion include, but are notlimited to, paraffin wax, carnauba wax, beeswax, candelilla wax, fruitwax, lanolin, shellac wax, bayberry wax, sugar cane wax,microcrystalline wax, ozocerite, ceresin, montan wax, and combinationsthereof. The hot melt glues include, but are not limited to, gluesincluding ethylene-vinyl acetate, polyethylene, polypropylene, apolyamide, or a polyester. One wax emulsion may be made up of 30%paraffin wax, 4% soy oil, 2% Span 60, 1% vitamin E, and distilled water.Another wax emulsion envisioned by the present invention includes 45%microcrystalline wax, 6% soy oil, 3% Span 60, 1% vitamin E, anddistilled water.

The semiochemical may be dissolved within the polymeric substrate,including within wax emulsions. As discussed above, the semiochemicalmay be a pheromone, or more specifically, may be an anti-aggregationpheromone. One particular anti-aggregation pheromone that may beutilized is verbenone. When verbenone is used, it may be present in thesystem in a range between about 0.001% by weight to about 60.0% byweight. More particularly, the verbenone may be present in the system inan amount of about 10% by weight.

Although the system may be utilized to control the population of a widerange of arthropods, this embodiment is also well suited for controllinginsect populations, and more particularly, coleopteran populations. Thesystem may be in a fluid form conducive to use within conventionalaerial spray equipment. The system may be prepared so as to release thesemiochemical over a long period of time, for example, over a 2-6 monthperiod.

Another embodiment of the present disclosure is directed toward a methodfor increasing the accessibility to control measures of an arthropodpopulation in a region. The arthropod is characterized by having asemiochemical-affected adult stage. In this method, a system isadministered to a region. As used herein, the region includes not onlythe orchard or plantation, but also the outer layers of individualtrees, surrounding ground cover, gardens, and individual plants. Thesystem administered to the region is of the type described above, thatis, the system is made up of a polymeric substrate and a semiochemical.The semiochemical is reactive upon the adult stage of the arthropod andis intermixed within the polymeric substrate.

The system may be administered by aerial spray equipment. Furthermore,the system may be administered by conventional aerial spray equipmentmounted on fixed-wing aircraft or rotorcraft. The system may also beadministered by terrestrial-based methods. For example, the system maybe administered by terrestrial-based spraying systems or by dispersingthe system by way of “paintballs.”

The method enables the novel effect of interfering with the behavior ofthe adult stage arthropod. As such, the method controls the populationof the arthropod by interacting with the adult stage of the arthropod,causing them to become more accessible to control measures including butnot limited to insecticides. Accordingly, the system may be administeredduring the immature stage of the arthropod and by being in the form of acontrolled-release, rainfast, substance, the system remains presentuntil the adult stage of the arthropod is reached. For example, in thecase of controlling CBB populations, the system may be administeredprior to coffee berry formation and the system may remain within theregion for at least six months thereby being present during the entirelife cycle of the CBB and being present to interfere with theadult-stage of the CBB. Accordingly, the semiochemical may repel anyadult-stage CBB present, thereby removing the CBB from the berry andincreasing accessibility to the CBB to better implement controlmeasures.

As discussed above, the semiochemical may be a pheromone, or moreparticularly may be an anti-aggregation pheromone. One pheromoneenvisioned for use in the method is verbenone. One particular embodimentof the method includes administering the system to the region such thateach hectare, or subparcel, of the region is administered 10 grams ofverbenone.

Another embodiment of the present disclosure is directed toward a methodof preparing a dispersible system for use in controlling the populationof arthropods. The steps of this method include providing a polymericsubstrate and adding a semiochemical to the polymeric substrate. Thesemiochemical is reactive upon an adult stage arthropod. Furthermore,the semiochemical may be a pheromone. In particular, the pheromone maybe an anti-aggregation pheromone. One particular pheromone that may beutilized is verbenone. The polymeric substrate may be a wax emulsion. Inthis embodiment, the wax emulsion may be formed by melting a wax, addingan oil, emulsifier, preservative, and water heated above the meltingtemperature of the wax to the melted wax to form a wax emulsion, andcooling the wax emulsion. Although many waxes are contemplated, thewaxes may include, but are not limited to, paraffin wax andmicrocrystalline wax. Additionally, the oil may be soy oil, theemulsifier may be Span 60, and the preservative may be vitamin E.

One embodiment of the present disclosure is directed toward methods fordislodging coffee berry borer from a coffee berry. The method involvesadministering a system on or in the vicinity of the coffee berry,wherein the system is made up of a polymeric or wax substrate and asemiochemical capable of affecting the coffee berry borer intermixedwithin the substrate. The substrate may be a wax emulsion, microspheres,a latex solution, hot melt glue, a resin, or plastic flakes. Thesemiochemical may be a pheremone, or more particularly ananti-aggregation pheremone. In particular, the anti-aggregationpheremone may be verbenone, methyl salicylate, an extract and infusionfrom the botanical orders Alismatales, Apiales, Arecales, Asparagales,Asterales, Brassicales, Canellales, Caryophyllales, Cycadales,Dipsacales, Ericales, Fabales, Gentianales, Lamiales, Laurales,Magnoliales, Malpighiales, Malvales, Myrtales, Pinales, Poales,Ranunculales, Rosales, Sapindales, Solanales, Theales or Zingiberales,or combinations of any thereof.

When the anti-aggregation pheremone is verbenone, it may be (−)Verbenone, (S) Verbenone, Levobernone, L-Verbenone and its respectivesisomers: (+) Verbenone, (R) Verbenone, D-Verbenone or any Verbenonecontaining isotopes of hydrogen (deuterium) and Carbon (13 Carbon) atany position in its structure. The verbenone may be present in thesystem in a range between about 0.001% by weight to about 60.0% byweight. In certain embodiments, the verbenone is present in the systemin an amount of about 10% by weight.

The system may be administered by either aerial spray equipment mountedon fixed-wing aircraft or rotorcraft or by a terrestrial-based sprayingsystem, and may be administered such that each hectare containing coffeeberries is administered 1-10,000 g of the semiochemical per hectare. Incertain embodiments, each hectare containing coffee berries isadministered 4,400 g of the semiochemical per hectare.

Another embodiment of the present disclosure is directed toward methodsof repelling arthropods in a region. The method features administering asystem to the region, wherein the system has a polymeric or waxsubstrate and an anti-aggregation pheremone intermixed within thesubstrate. The substrate may be a wax emulsion, microspheres, a latexsolution, hot melt glue, a resin, or plastic flakes. Theanti-aggregation pheremone may be verbenone, methyl salicylate, anextract and infusion from the botanical orders Alismatales, Apiales,Arecales, Asparagales, Asterales, Brassicales, Canellales,Caryophyllales, Cycadales, Dipsacales, Ericales, Fabales, Gentianales,Lamiales, Laurales, Magnoliales, Malpighiales, Malvales, Myrtales,Pinales, Poales, Ranunculales, Rosales, Sapindales, Solanales, Thealesor Zingiberales, or combinations of any thereof.

The method may be used to repel insects, and more particularlycoleopterans. Indeed, it is envisioned that the method may be used torepel coleopteran from the genera: Dendroctonus spp., Rhynchophorusspp., Hypothenemus spp., Xylosandrus spp., Cosmopolites spp.,Alphitobius spp., Sphenophorus spp., and Anthonomus spp. The method isenvisioned to particularly work to repel the coffee berry borer. In thatregard, further testing has shown that a combination of verbenone andwintergreen oil is particularly beneficial in repelling a multitude ofcoleopterans. Wintergreen oil, and its main constituent, methylsalicylate, can be prepared as described in U.S. patent application Ser.No. 15/143,097, titled Methods and Compositions for ControllingLeaf-Cutting Ant Populations, filed on Apr. 29, 2016, the entirety ofwhich is incorporated by reference herein. In particular, wintergreenoil is derived from a group of plants of the genus Gaultheria (multiplecommon names, including American mountain tea, boxberry, creepingwintergreen, spice berry, and hillberry, among others). The term“wintergreen” is a somewhat antiquated reference to the ability of theseplants to sustain photosynthetic activity throughout the winter; theterm “evergreen” is now more commonly used to describe thischaracteristic. Wintergreen oil is derived from Gaultheria plants bysteam distillation of their leaves, following maceration in warm water.This process produces an enzymatic action by a glycoside within theleaves, yielding the main constituent of wintergreen oil, methylsalicylate. The oil itself is a pale yellow or pinkish fluid that isstrongly aromatic, with a sweet woody odor. The components ofwintergreen oil are: methyl salicylate (˜98%), α-pinene, myrcene,delta-3-carene, limonene, 3,7-guaiadiene, and delta-cadinene, whichgives wintergreen plants a distinctive “medicinal” smell wheneverbruised.

Yet another embodiment of the present disclosure is directed towardsystems for repelling coffee berry borer. The system includes apolymeric or wax substrate and an anti-aggregation pheremone intermixedwithin the substrate. The substrate may be a wax emulsion, microspheres,a latex solution, hot melt glue, a resin, or plastic flakes. Theanti-aggregation pheremone may be verbenone, methyl salicylate, anextract and infusion from the botanical orders Alismatales, Apiales,Arecales, Asparagales, Asterales, Brassicales, Canellales,Caryophyllales, Cycadales, Dipsacales, Ericales, Fabales, Gentianales,Lamiales, Laurales, Magnoliales, Malpighiales, Malvales, Myrtales,Pinales, Poales, Ranunculales, Rosales, Sapindales, Solanales, Thealesor Zingiberales, or any combinations thereof.

The system may further include an insect toxicant. The substrate may bea wax emulsion formed by melting a wax and adding to the melted wax, anoil, an emulsifier, a preservative, and water heated above the meltingtemperature of the wax to form the wax emulsion. The wax emulsion isthen cooled. In one particular embodiment, the system has the formulashown in Table 1 below.

TABLE 1 water 5%-60% by weight; petrolatum 2%-20% by weight; soybean oil5%-80% by weight; sorbitan monostearate 0.1%-10% by weight; fumed silica0.5%-10% by weight; wax 1%-50% by weight; activated charcoal 0.1%-5% byweight; verbenone 1%-60% by weight; and methyl salicylate 1%-60% byweight.

Alternatively, the wax emulsion may be made up of a wax carrier selectedfrom paraffin wax, carnauba wax, beeswax, candelilla wax, fruit wax,lanolin, shellac wax, bayberry wax, sugar cane wax, microcrystallinewax, ozocerite, ceresin, montan wax, or combinations thereof. Oneparticular embodiment has a wax emulsion with the components listed inTable 2.

TABLE 2 30% by weight paraffin wax; 4% by weight soy oil; 2% by weightsorbitan monostearate 1% by weight vitamin E; and 58% by weightdistilled water. The system of claim 3, wherein the wax emulsioncomprises: 45% by weight microcrystalline wax; 6% by weight soy oil; 3%by weight sorbitan monostearate; 1% by weight vitamin E; and 40% byweight distilled water.

The substrate may alternatively be a hot melt glue, in particular it maybe a polymer selected from ethylene-vinyl acetate, polyethylene,polypropylene, a polyamide, or a polyester.

The formulations described above may further include water, wherein thewax carrier is in a fluid suspension of the water. The fluid suspensionmay be an emulsion. The formulations may further include one or moreadditives selected from lipids, emulsifiers, plasticizers, UV blockersand absorbers, antimicrobials, antioxidants and volatility suppressants.These additives, if present, may be in an amount from about 0.001% toabout 20% by weight. The emulsifiers may be lipids, soy oil, lecithins,modified lecithins, monoglycerides, diglyceridies, sorbitans, fattyacids, or combinations thereof. The emulsion may be sprayable,squirtable or spreadable.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features and advantages of the various embodimentsdisclosed herein will be better understood with respect to the followingdescription and drawings, in which like numbers refer to like partsthroughout, and in which:

FIG. 1 is a graph showing coffee berry infestation between controlgroups and those being treated with a composition as described herein;and

FIG. 2 is a graph showing coffee berry infestation as described in FIG.1 over a period of time.

DETAILED DESCRIPTION

The detailed description set forth below is intended as a description ofthe presently preferred embodiment of the invention, and is not intendedto represent the only form in which the present invention may beconstructed or utilized. The description sets forth the functions andsequences of steps for constructing and operating the invention. It isto be understood, however, that the same or equivalent functions andsequences may be accomplished by different embodiments and that they arealso intended to be encompassed within the scope of the invention.

Semiochemical formulations should exhibit a zero-order-release rate andsustain release levels above a certain threshold for a long period oftime, wherein release levels below threshold would only have anegligible disruptive effect on the behavior of the target insect. Witha couple of exceptions, when formulated with less than 10% activeingredient (Al) Specialized Pheromone and Lure Application Technology(SPLAT) consistently provides a near zero-order-release rate of thesemiochemical, with negligible flash-off around the time of application.

The initial research and development that culminated into the existingSPLAT technology was done using Grapholita molesta, the Oriental fruitmoth (OFM), a serious pest of apples worldwide, as the model insect.When formulated with less than 10% OFM pheromone, SPLAT consistentlyprovided a near zero-order-release rate with negligible flash-off. Fieldtrials in large commercial apple operations in South America indicatedthat SPLAT formulations containing 15 g pheromone per acre sustainednearly complete trap shutdown for over 180 days, which actuallytranslated in a significant reduction of OFM fruit damage, as comparedto that found in the grower's traditional chemical control. Analysis ofthe field “aged” SPLAT indicated that there were different levels ofpheromone remaining in the point source of SPLAT by the end of the 180day trial period, and it was related to the position of the dollop inthe canopy (receiving more or less solar incidence) as well as theactual size of the dollop analyzed: pheromone in microdollops wasundetectable, whereas 1 g dollops still contained 5-10% and 5 to 10 gdollops retained between 10 and 25% AI.

In the case of CBB, the present disclosure envisions a SPLAT andpheromone based repellent formulation that provides acceptable long-termrepellency while minimizing any potential deleterious impacts. Theapplication of SPLAT and verbenone can occur at the onset of coffeeberry formation and by remaining in place emitting pheromone would repelexisting and emerging adults from further damaging developing coffeeberries.

This novel formulation, a SPLAT and verbenone formulation that isrepellent, can be applied using conventional defensive applicationapparatus, will last for 6 months or longer in the field, releasesnearly all the pheromone it contains, and is biodegradable and safe.

This formulation provides coffee farmers with a functional system andsubstantial savings. The cost of SPLAT with 10 g verbenone/acre will beabout $30 dollars per acre, which represents a savings of $30-60 peracre just considering the cost of BOTANIGARD®ES. Now, if thisformulation also substitutes the customary two treatments ofBOTANIGARD®ES at a cost of $60-90/acre, then the savings will be$60-120/acre.

Our calculation of the savings is very conservative. It does not take inaccount the additional savings realized because of the simplification ofthe current cultural control CBB operation, a formulation with such along life that it allows for a single application for the entire CBBcycle. More savings will be achieved by elimination of crisis situationsand their associated cost. Furthermore, we believe we overestimate thecost of manufacturing SPLAT. It is probably high because it is based onthe price we currently pay our suppliers, which do not reflect thediscounts available from commercial suppliers when we purchase the rawmaterials in large bulk quantities. Furthermore the efficiency of theproduction line increases with the larger volumes, needing less workerhours per volume of SPLAT produced. These additional savings have notbeen accounted for.

Because of its repellent action, SPLAT and verbenone may providepopulation protection not only in areas of existing populations but alsoareas with low and no population densities, such as newly establishedplantations, providing a huge benefit to the coffee industry andpopulation in general.

It is believed that the ideal verbenone formulation should be appliedusing conventional spray equipment, have a duration of at least two tosix months, stick to the foliage where it lands, quickly acquirerain-fast qualities, protect the pheromone from degradation, worksynergistically with adulticides so that it can possibly control CBBpopulations at low as well as at high densities, be biodegradable, ifpossible organic, not damage private property, and last but not leastthe formulation should be inexpensive for its adoption to be not onlytechnically, but also economically feasible.

The present invention is expected to meet all of the desired factors byproviding an optimal semiochemical solution for the effective managementof CBB independent of population density. The overall objective of thisinvention is to provide effective season-long field control of CBBpopulations using a flowable wax emulsion system (SPLAT) that deliversthe anti-aggregation pheromone verbenone. We formulated SPLAT andverbenone using flowable wax emulsions of different characteristics inorder to determine in the lab the emission rate and stability of thepheromone. Two of the optimized formulations were submitted tolaboratory and field trials to which we added high CBB pressure. Fieldaged samples of the SPLAT formulations were analytically quantified todetermine residual stability and effectiveness of pheromone components.

Some of the goals of the invention include: 1) a formulation having aduration of six months while being protected from degradation whiledispensing verbenone; 2) a formulation that works synergistically withkilling agents; 3) a formulation of SPLAT that works with conventionalaerial spray equipment, sticks to foliage and quickly becomes rain-fast;and 4) a formulation that controls CBB populations at low as well ashigh densities. However, it is contemplated that in some embodiments ofthe invention only some, or even none of the goals may be achieved.

There are no verbenone formulations in the market today that have highlongevity and that can be applied using conventional spray equipment.The present invention is an optimal semiochemical solution for theeffective management of CBB independent of population density. Thepresent invention is effective in season-long field control of CBBpopulations using a flowable wax emulsion system (SPLAT) that deliversthe anti-aggregation pheromone verbenone. The release rate of verbenonefrom SPLAT, 10% of pheromone in the SPLAT composition, applied in thefield as point sources at three sizes, 1 g, 7 g and 17 g each wasmeasured. The SPLAT formulations retained and continued emittingpheromone for 120 days, with the formulations still retaining 30%-80% ofits pheromone (depending on dose and point source size). This suggeststhat these formulations would probably last another 30 to 60 days in thefield (depending on dose and point source size). This indicates that ifthe formulation is sprayed in the field at the time of coffee berrydevelopment, it will last through harvest. This allows for the firsttime a formulation that can be sprayed to control CBB through theemission of effective rates of pheromone during adult flight.

By targeting CBB with an organic, safe, and effective formulation,non-target organisms will be minimally affected by any extra insecticideuse, so insect species diversity will be retained where thesemiochemical formulation is used. This in turn will conserve vertebrateanimals that depend upon insects for their diet.

In one embodiment of SPLAT and verbenone the pheromone lasts for thewhole season, but just for the coffee berry production season, notlonger, to avoid unwanted residual problems and exposure to non-targetspecies. The systems and methods of the present invention may beutilized for a plethora of other pests and invasive species (e.g.,Mountain Pine Beetle, Southern Pine Beetle, Western Pine Beetle andothers).

Our testing involved application of approximately 0.2 g of SPLAT Verb inapproximately a 6-inch petri dish. (SPLAT Verb is loaded with 10%verbenone by weight.) This area would correspond to 0.0000045ac of land;therefore, our field application rate would be approximately 44 kg ofSPLAT Verb (4,400 g of verbenone) per acre. Coffee berries, commonlyused to rear CBB, were placed in the petri dish. The 0.2 g of SPLAT Verbwas introduced into the sealed petri dish. We then went on to measurethe times when CBB dislodged or exited the berry. The number dislodgedis a summation of the total dislodged at that point in time. Our data(as shown in Table 3) verifies that CBB exited their preferred host and,after dislodging, maintained high activity levels without re-enteringthe berries. This data serves as the first confirmation of verbenone'srepellant activity.

TABLE 3 Number Activity Level Time Dislodged Outside of Berry  4 min 51sec 2 High  7 min 17 sec 3 High 15 min 32 sec 4 High 19 min 25 sec 6High 22 min 21 sec 7 High 28 min 38 sec 8 High 32 min 32 sec 9 HighTotal in 37 min 9 High

Alcohol is a strong CBB attractant. Further testing of SPLAT Verb shows(as can be seen in FIGS. 1 and 2) that the addition of verbenone reducesthe level of attraction of CBB even to traps with alcohol to levelsbelow the control (trap without attractant or any other semiochemical).

The SPLAT and verbenone pheromone release formulation, a novel,amorphous, flowable emulsion can be applied as microliter point sourcesall the way to dollops of tens of grams. The SPLAT wax dispenserformulations of this invention belong to a “matrix-type” or “monolithic”category of controlled-release devices. These “matrix-type” or“monolithic” dispensers are defined as devices where the activeingredient is dispersed or dissolved in a polymer matrix. Release of theactive ingredient from a monolithic device occurs by diffusion and canbe described macroscopically by Fick's Law. Fick's law states that themovement of a molecule by diffusion is directly proportional to theconcentration of that molecule in a system. Microscopically, if wefollow the movement of a molecule of an active agent through a matrix,this molecule can begin its journey in one of two ways. If it isdispersed in the matrix, it begins its journey by dissociating fromother molecules in its crystal cell and solubilizing into the polymerphase. If it is dissolved in the matrix, then this step is bypassed. Themolecule then diffuses through amorphous regions in the matrix thatcomprise the free volume of the system. The molecule can move throughthe matrix in one of two ways as well. If it is very small compared tothe size of the amorphous spaces in the matrix, then it will diffusethrough the matrix by moving from one such space to another. If it isvery large compared to the size of those spaces, then segments of thepolymer comprising the matrix will have to be rearranged for diffusionof the active agent molecule to occur. Crystalline regions in the matrixare virtually impermeable to molecules of the active agent. Uponreaching the surface of the matrix, it will be released into theenvironment.

A series of factors influences the rate of release of an active agentfrom a monolithic device and includes properties of the matrix materialas well as properties of the active agent. The temperature of the matrixinfluences release of the active agent; at higher temperatures the freevolume is increased, and diffusion occurs faster. At lower temperatures,the free volume is decreased, and diffusion is slower. The thermalhistory of a polymer can also increase or decrease the free volume ofthe system and lead to changes in the diffusional rate of an activeagent.

The property of the active agent having the greatest influence on itsrelease rate is its molecular weight. Generally, larger molecules takemore time to make their way through the free space of a matrix.Branching in a molecule can also decrease its rate of diffusion througha matrix. The partition coefficient of the active agent between thematrix and the environment can also influence the release rate of thatagent. If the agent readily partitions to the environment, then its rateof release will be diffusion-controlled and first order. If, however,partitioning of the active agent to the environment is relatively slow,then its partition coefficient will determine its release rate from thematrix and the device will exhibit zero order release kinetics. Thepartitioning of the active agent to the environment is a function of thesolubility of the active agent in the matrix; compounds more soluble inthe matrix partition to the environment more slowly. SPLAT paraffinemulsions in a field environment exhibit diffusion-controlled release.The surface area of the device also influences its release rate.Paraffin dispensers with larger surface areas release active agent atfaster rates.

The release rate of a SPLAT formulation containing a fixed amount ofsemiochemical can be modulated simply by changing a few of parameters ofthe formulation, which includes the type of components used (e.g. thewax composition, the emulsifiers used), their proportion in theformulation (e.g., percentage of water, oil or wax content), the stagein the manufacturing the different components are added, the rheology,and finally the characteristics of application of the SPLAT in the field(e.g., applied as microdollops of 1-10 ug each or large dollops of 10 geach).

The result is a semiochemical formulation that is extremely malleableand that fits many needs and uses that cannot be supported by any othercommercial formulation present in the market.

A 30% paraffin wax emulsion was made consisting of 30% paraffin wax(Gulf Wax, Royal Oak Sales, Inc., Roswell, Ga.), 4% soy oil (SpectrumNaturals, Inc., Petaluma, Calif.), 2% Span 60 (Sorbitan monostearate,Sigma-Aldrich Co., St. Louis, Mo.), 1% vitamin E α-tocopherol, SigmaChemical Co., St. Louis, Mo.), and 58% distilled water. A 45%microcrystalline wax emulsion consisting of 45% microcrystalline wax(Blended Waxes, Inc., Oshkosh, Wis.), 6% soy oil, 3% Span 60, 1% vitaminE, and 40% distilled water was also made.

The wax is melted (paraffin: 60-65° C.; microcrystalline: 78-80° C.) andwater heated above the melting temperature of the wax (paraffinemulsion: 65-70° C.; microcrystalline emulsion: 78-88° C.). The soy oil,Span 60, and vitamin E are added to the melted wax and thoroughly mixed,followed by the addition of the hot water. This mixture is then pouredinto a industrial laboratory blender. The emulsion is immediatelyblended, then placed in a cold water bath, and mixed every 15 minutesuntil the solution had cooled to 25-30° C. when it is placed in aplastic bucket and stored until use.

Just prior to use, 0.03% (3 g), 1.0% (10 g) and 3.0% (30 g) by weight ofemulsion of racemic verbenone is thoroughly mixed into the emulsionusing a high sheer lab mixer.

Preliminary work with generic SPLAT formulations containing 3% racemicverbenone using flow cells indicates that it releases pheromone at avery constant level for long periods. To show the release of AIsemiochemicals within SPLAT, we collected the effluvia from 5 g SPLAT GM3%, containing 150 mg disparlure (AI) and found that it releaseddisparlure at a rate of 44.06±13.08 ug/day for over ca. 170 days. As acomparison, 5 g of Disrupt II (a prior art formulation), containing 894mg disparlure emit 51.45±2.33 ug/day. It is important to realize thatalthough both flow chambers had 5 grams of formulation, Disrupt IIactually had six times more pheromone than SPLAT while it released only15% more pheromone than SPLAT, a difference that probably has nobiological effect to speak of. These results suggest that SPLAT is amuch more efficient formulation than those known in the prior art;actually six times more efficient. As such, one would need to apply sixtimes less AI using SPLAT than if using prior art formulations. Usuallythe most expensive component of a mating disruption formulation is theactive ingredient. Accordingly, SPLAT is believed to be substantiallyless expensive than the formulations of the prior art.

With SPLAT, one can change the consistency of the emulsion by changingthe proportion of components, or by changing the rheology of the mixingof the components. The word “rheology” normally refers to the flow anddeformation of “non-classical” materials such as rubber, moltenplastics, polymer solutions, slurries and pastes, electrorheologicalfluids, blood, muscle, composites, soils, and paints. These materialscan exhibit varied and striking external and internal structures due totheir rheological properties that classical fluid mechanics andelasticity cannot describe. Our experience is that the SPLAT formulawith 45% microcrystalline wax emulsion (45% microcrystalline wax, 6% soyoil, 3% Span 60, 1% vitamin E, and 40% distilled water) can bemechanically applied and that it adheres quickly to the vegetation, andas long as it has a couple hours to settle, it becomes rain fast.

Specialized Pheromone & Lure Application Technology (SPLAT) is a basematrix formulation of biologically inert materials used to control therelease of semiochemicals and/or odors with or without pesticides.Extensive research on SPLAT using a variety of lures demonstrates thatthis matrix emits semiochemicals at effective pest suppression levelsfor a time interval ranging from 2-16 weeks. Having a wide range ofviscosities and application methods (e.g. applicator sprays, aerialapplicator sprays, caulking gun type tubes, etc.), SPLAT increasesproductivity by mechanizing the application of pheromone dispensingpoints. The amorphous and flowable quality of this highly adaptableproduct allows for an easy transition from small-scale manualapplications to large-scale mechanical applications.

Additional experimentation as described below shows that thecompositions and methods described herein function with numerouscoleopterans. Testing was done to evaluate the effect of SPLAT plusVerbenone (+) and Wintergreen Oil on the Lesser Mealworm, Alphitobiusdiaperinus. To do so, the composition was placed in traps containing asubstrate attractant to see whether there would be a reduction incaptured insects. Four repetitions were used for each treatment. Toavoid any interference between the treatments, a minimum distance of 25meters was used to separate each trap. The treatments were prepared inthree formats: 01 had the trap with poultry litter, 02 had the trap withpoultry litter and the SPLAT plus Verbenone (+) and Wintergreen Oilcomposition, and 03 was merely the trap. The number of captured insectsafter two weeks were determined. The results are shown in Table 4 below.As such, it can be seen that the presence of SPLAT with Verbenone(+)+Wintergreen Oil reduce the number of insects captured in traps withan attractant source (poultry litter), showing a repellent effect forthe species Alphitobius diaperinus.

TABLE 4 Treatment R1 R2 R3 R4 Average 01 48 76 29 66 54.7 02 3 0 5 2 2.503 0 0 0 0 0

Testing was done to evaluate the effect of SPLAT plus Verbenone (+) andWintergreen Oil on the behavior of the Banana root borer, Cosmopolitessordidus. To do so, the composition was placed in traps containing anattractant source (pheromone traps) to see whether there would be areduction in captured insects. Four repetitions were used for eachtreatment. To avoid any interference between the treatments, a minimumdistance of 25 meters was used to separate each trap. The treatmentswere prepared in three formats: 01 had the trap with pheromone, 02 hadthe trap with pheromone and the SPLAT plus Verbenone (+) and WintergreenOil composition, and 03 was merely the trap. The number of capturedinsects after four weeks were determined. The results are shown in Table5 below. As such, it can be seen that the presence of SPLAT withVerbenone (+)+Wintergreen Oil reduce the number of insects captured intraps with an attractant source (pheromone), showing a repellent effectfor the species Cosmopolites sordidus.

TABLE 5 Treatment R1 R2 R3 R4 Average 01 18 22 38 24 25.5 02 1 0 2 00.75 03 0 0 0 1 0.25

Testing was done to evaluate the effect of SPLAT plus Verbenone (+) andWintergreen Oil on the behavior of the Sugarcane weevil, Sphenophoruslevis. To do so, the composition was placed in traps containing anattractant source (fermented sugarcane) to see whether there would be areduction in captured insects. Four repetitions were used for eachtreatment. To avoid any interference between the treatments, a minimumdistance of 25 meters was used to separate each trap. The treatmentswere prepared in three formats: 01 had the trap with fermentedsugarcane, 02 had the trap with fermented sugarcane and the SPLAT plusVerbenone (+) and Wintergreen Oil composition, and 03 was merely thetrap. The number of captured insects after six weeks were determined.The results are shown in Table 6 below. As such, it can be seen that thepresence of SPLAT with Verbenone (+)+Wintergreen Oil reduce the numberof insects captured in traps with an attractant source (fermentedsugarcane), showing a repellent effect for the species Sphenophoruslevis.

TABLE 6 Treatment R1 R2 R3 R4 Average 01 11 7 26 12 14 02 1 0 0 0 0.2503 0 0 0 0 0

Testing was done to evaluate the effect of SPLAT plus Verbenone (+) andWintergreen Oil on the behavior of the Boll Weevil, Anthonomus gradis.To do so, an “X” olfactometer was used to realize the choice tests.Twenty insects were used per treatment. Namely, two of the fourextremities received a control treatment (blank SPLAT) and the other twothe specific composition. The treatments were prepared in two formats:01 the control, made up of Blank SPLAT and a pheromone (grandlure) and02 the test, made up of Blank SPLAT, pheromone (grandlure) with theVerbenone (+) and Wintergreen Oil composition. The number of insectsthat moved into each treatment extremity of the olfactometer wasrecorded. The results are shown in Table 7 below. As such, it can beseen that he presence of SPLAT with Verbenone (+)+Wintergreen Oil reducechoice number of insects when used in combination with an attractantsource (pheromone), showing a repellent effect for the speciesAnthonomus grandis.

TABLE 7 Control Pheromone (Blank SPLAT) (Grandlure) Treatment 01 02 18Pheromone (Grandlure) + Control SPLAT plus Verbenone (+) + (Blank SPLAT)Wintergreen Oil Treatment 02 16 04

The above description is given by way of example, and not limitation.Given the above disclosure, one skilled in the art could devisevariations that are within the scope and spirit of the inventiondisclosed herein, including various ways of administering the verbenone.Further, the various features of the embodiments disclosed herein can beused alone, or in varying combinations with each other and are notintended to be limited to the specific combination described herein.Thus, the scope of the claims is not to be limited by the illustratedembodiments.

What is claimed is:
 1. A system for repelling coleopterans, the systemcomprising: a polymeric or wax substrate selected from the groupconsisting of a wax emulsion, microspheres, a latex solution, hot meltglue, a resin, and plastic flakes; and an anti-aggregation pheremoneintermixed within the substrate, wherein the anti-aggregation pheremoneis selected from the group consisting of verbenone, methyl salicylate,wintergreen oil extracted from the leaves of a plant within theGaultheria genus, an extract and infusion from the botanical ordersAlismatales, Apiales, Arecales, Asparagales, Asterales, Brassicales,Canellales, Caryophyllales, Cycadales, Dipsacales, Ericales, Fabales,Gentianales, Lamiales, Laurales, Magnoliales, Malpighiales, Malvales,Myrtales, Pinales, Poales, Ranunculales, Rosales, Sapindales, Solanales,Theales or Zingiberales, and combinations thereof.
 2. The system ofclaim 1, further comprising an insect toxicant.
 3. The system of claim1, wherein the substrate is a wax emulsion formed by: melting a wax; andadding to the melted wax, an oil, an emulsifier, a preservative, andwater heated above the melting temperature of the wax to form the waxemulsion.
 4. The system of claim 1, wherein the anti-aggregationpheremone is a combination of verbenone and methyl salicylate.
 5. Thesystem of claim 1, wherein the anti-aggregation pheremone is acombination of verbenone and wintergreen oil extracted from the leavesof a plant within the Gaultheria genus.
 6. The system of claim 1,wherein the anti-aggregation pheremone is a combination of verbenone,methyl salycylate, and wintergreen oil extracted from the leaves of aplant within the Gaultheria genus.
 7. The system of claim 1, wherein thecoleopteran is from the genera Dendroctonus spp., Rhynchophorus spp.,Hypothenemus spp., Xylosandrus spp., Cosmopolites spp., Alphitobiusspp., Sphenophorus spp., and Anthonomus spp.
 8. The system of claim 7,wherein the coleopteran is the coffee berry borer.
 9. The system ofclaim 7, wherein the coleopteran is Cosmopolites sordidus.
 10. Thesystem of claim 7, wherein the coleopteran is Alphitobius diaperinus.11. The system of claim 7, wherein the coleopteran is Sphenophoruslevis.
 12. The system of claim 7, wherein the coleopteran is Anthonomusgrandis.
 13. A method of repelling coleopterans in a region, the methodcomprising administering a system to the region, the system comprising:a polymeric or wax substrate selected from the group consisting of a waxemulsion, microspheres, a latex solution, hot melt glue, a resin, andplastic flakes; and an anti-aggregation pheremone intermixed within thesubstrate, wherein the anti-aggregation pheremone is selected from thegroup consisting of verbenone, methyl salicylate, wintergreen oilextracted from the leaves of a plant within the Gaultheria genus, anextract and infusion from the botanical orders Alismatales, Apiales,Arecales, Asparagales, Asterales, Brassicales, Canellales,Caryophyllales, Cycadales, Dipsacales, Ericales, Fabales, Gentianales,Lamiales, Laurales, Magnoliales, Malpighiales, Malvales, Myrtales,Pinales, Poales, Ranunculales, Rosales, Sapindales, Solanales, Thealesor Zingiberales, and combinations thereof.
 14. The method of claim 13,wherein the anti-aggregation pheremone is a combination of verbenone andmethyl salicylate.
 15. The method of claim 13, wherein theanti-aggregation pheremone is a combination of verbenone and wintergreenoil extracted from the leaves of a plant within the Gaultheria genus.16. The method of claim 13, wherein the anti-aggregation pheremone is acombination of verbenone, methyl salycylate, and wintergreen oilextracted from the leaves of a plant within the Gaultheria genus. 17.The method of claim 13, wherein the coleopteran is from the generaDendroctonus spp., Rhynchophorus spp., Hypothenemus spp., Xylosandrusspp., Cosmopolites spp., Alphitobius spp., Sphenophorus spp., andAnthonomus spp.
 18. A method for dislodging coffee berry borer from acoffee berry, the method comprising administering a system on or in thevicinity of the coffee berry, the system comprising: a polymeric or waxsubstrate selected from the group consisting of a wax emulsion,microspheres, a latex solution, hot melt glue, a resin, and plasticflakes; and an anti-aggregation pheremone intermixed within thesubstrate, wherein the anti-aggregation pheremone is selected from thegroup consisting of verbenone, methyl salicylate, wintergreen oilextracted from the leaves of a plant within the Gaultheria genus, anextract and infusion from the botanical orders Alismatales, Apiales,Arecales, Asparagales, Asterales, Brassicales, Canellales,Caryophyllales, Cycadales, Dipsacales, Ericales, Fabales, Gentianales,Lamiales, Laurales, Magnoliales, Malpighiales, Malvales, Myrtales,Pinales, Poales, Ranunculales, Rosales, Sapindales, Solanales, Thealesor Zingiberales, and combinations thereof.
 19. The method of claim 18,wherein the anti-aggregation pheremone is a combination of verbenone andmethyl salicylate.
 20. The method of claim 18, wherein theanti-aggregation pheremone is a combination of verbenone and wintergreenoil extracted from the leaves of a plant within the Gaultheria genus.21. The method of claim 18, wherein the anti-aggregation pheremone is acombination of verbenone, methyl salycylate, and wintergreen oilextracted from the leaves of a plant within the Gaultheria genus.